Potential candidate cells for constructing tissue-engineered lacrimal duct epithelium: a histological and cytological study in rabbits

• Published in: Journal of Zhejiang University. B. Science Vol. 16; no. 11; pp. 904 - 913
• Main Authors: Xie, Chen, Li, Xiu-yi, Cui, Hong-guang
• Format: Journal Article
• Language: English
• Published: Hangzhou Zhejiang University Press 01.11.2015; Springer Nature B.V
• Subjects: Animals; Biomedical and Life Sciences; Biomedicine; Cell Proliferation; Cells, Cultured; Cellular structure; Conjunctiva; Construction engineering; Cytokeratin; Diseases; Electron microscopy; Emission; Epithelial cells; Epithelial Cells - cytology; Epithelial Cells - ultrastructure; Epithelium; Goblet cells; Immunohistochemistry; In vitro methods and tests; Lacrimal Apparatus - cytology; Lacrimal Apparatus - ultrastructure; Light microscopy; Male; Markers; Microscopy, Electrochemical, Scanning; Physical characteristics; Proliferating cell nuclear antigen; Rabbits; Scanning electron microscopy; Staining; Tissue Engineering; Transmission electron microscopy; Lacrimal duct epithelial cells; Cytokeratin; 结膜上皮细胞; Tissue engineering; 黏蛋白; R777.2; 泪道上皮细胞; Q813; 组织工程; 细胞角蛋白; Conjunctival epithelial cells; Mucin; 人增殖细胞核核抗原; Proliferating cell nuclear antigen
• ISSN: 1673-1581; 1862-1783
• DOI: 10.1631/jzus.B1500113

Objective Injury and deficiency of the lacrimal duct epithelium (LDE) can lead to a variety of lacrimal diseases. The purpose of this study was to characterize potential candidate cells for constructing a tissue-engineered LDE. Method Different areas of the conjunctiva and lacrimal duct tissue were removed from male adult New Zealand white rabbits for histological evaluation. Hematoxylin and eosin staining and immunohistochemical staining of cytokeratin AE1+AE3, cytokeratin 4, Ki-67, and MUC5AC were observed by light microscopy. The surface morphologies of different epithelial tissues and cellular structures were examined using field-emission scanning electron microscopy and transmission electron microscopy. Epithelial cells were isolated from tissues and identified by specific markers. In vitro , proliferative ability and Western blot analyses of the proliferating cell nuclear antigen (PCNA) of different epithelial cells cultured in identical environments were investigated and compared. Results Histologically, the epithelial specific markers, cytokeratin AE1+AE3 and cytokeratin 4, were expressed in the conjunctiva epithelium and the LDE. Notably, highly proliferative cells stained with Ki-67 were concentrated under the epithelium in a dome structure of the posterior palpebral conjunctiva. Differentiated goblet cells were also found to a lesser extent in this region. Primary palpebral and fornical conjunctival epithelial cells (PFCECs), bulbar conjunctival epithelial cells (BCECs), and lacrimal duct epithelial cells (LDECs) were successfully separated from tissues. In vitro , rabbit PFCECs and LDECs grew faster and expressed more PCNA than BCECs. Conclusions PFCECs are anatomically similar to LDECs. They also have similar morphological characteristics, immune phenotypes, and proliferation features. PFCECs are therefore potential candidate cells to replace LDECs in tissue engineering to treat lacrimal duct diseases.